KR20070014291A - Ginseng polysaccharides composition having the use of stabilization and propagation of lactic acid bacteria - Google Patents

Abstract

A polysaccharide composition of ginseng containing one or more components of ginseng, red ginseng or high temperature treated ginseng is provided. The composition has effects of stabilizing and proliferating lactic acid bacteria and suppressing the production of an enzyme that seems to cause cancer such as large intestinal cancer. The polysaccharide composition of ginseng contains 1 to 99.9% by weight of one or more components of ginseng, red ginseng or high temperature treated ginseng, based on the total weight of the composition. The polysaccharide contains 40 to 50% by weight of a hexose including glucose or galactose, 30 to 50% by weight of an acidic polysaccharide including galacturonic acid or rhamnose or 3 to 6% by weight of protein including histidine, arginine, aspartic acid, threonine, serine or glutamic acid. The polysaccharide is prepared by extracting ginseng, red ginseng or high temperature treated ginseng with 1 to 20 the volume of lower alcohol such as ethanol at 4 to 100deg.C for 0.5hr to 2 days, filtering and then extracting the residue with 1 to 20 the volume of distilled water at 4 to 150deg.C for 1 to 24hr.

Description

Ginseng polysaccharide composition having stabilization and lactic acid bacteria growth effect of lactic acid bacteria {GINSENG POLYSACCHARIDES COMPOSITION HAVING THE USE OF STABILIZATION AND PROPAGATION OF LACTIC ACID BACTERIA}

Figure 1 shows the elution pattern of the red ginseng polysaccharide fraction in the Sephadex G-75 column.

The present invention relates to a composition containing ginseng polysaccharide as an active ingredient. In addition, the present invention relates to a composition having a lactic acid bacteria freeze-drying protective effect, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect and the production inhibitory effect of causative enzymes such as colon cancer.

Ginseng is a perennial root of the genus Ogasus ginseng by plant taxonomy. About 11 species of ginseng are known on the earth. For example, Ginseng is native to the Far East of Asia (33-48 ° north: Korea, Northern Manchuria, and Russia) and has very good efficacy. Korean ginseng (Panax ginseng CAMeyer); American ginseng (Panax quinquefolium L.) growing and growing in the United States and Canada; Panax notoginseng F.H.Chen, wild or cultivated in southwestern Guangxi province from southeastern Yunnan Province of China; And Panax japonicus C.A.Meyer, which is distributed in Japan, southwest China, and Nepal.

Ginseng is not only stored as a commodity in the new agricultural plant, but has been used as a valuable medicine since ancient times. Many pharmacological experiments to date have revealed that ginseng strengthens the nonspecific resistance of the body to stress and has an acidic action. In addition, it has been shown to improve hypertension, enhance insulin action, hypoglycemic effect in alloxan diabetic mice, liver RNA synthesis in rats, protein synthesis, sugar and lipid metabolism promoting effect, and anticancer effect.

The ginseng has been used for various diseases such as psychiatric diseases, nervous system diseases and diabetes in the form of herbal medicine mainly in Asian countries such as Korea, China, Japan, and saponin, which is a major component of the ginseng, is tonic It is known to have effects on stiffness, sedation, and antihypertension.

Currently, the use of ginseng is white ginseng, dried ginseng as it is grown and collected at room temperature, red ginseng prepared by heating at 98-100 ° C, or heat-treated ginseng prepared by heating at 120-180 ° C. , Black ginseng, etc.). The components of ginseng are polysaccharides, ginsenosides, polyacetylene, nucleic acid base glycosides (nucleosides), alkaloids, phenolic compounds, proteins, amino acids, fragrances that are also present in ginseng. Etc. are revealed. Ginseng polysaccharides were further classified into acidic, neutral and alkaline polysaccharides, and ginsenosides, which are specific to ginseng, were isolated from 34 species (22 white ginseng and 30 red ginseng), and recently, compound K (20-O-β). Ginseng saponin metabolites such as -D-glucopyranosyl-20 (S) -protopanaxadiol) have been reported separately. As currently known, the anti-cancer effect, anti-diabetic effect, and immuno-enhancing effect among the pharmacological effects of ginseng were found to be due to the polysaccharide and saponin of ginseng.

About 300 kinds of bacteria and more than 100 trillion bacteria live in the intestines of humans, and have a significant impact on human health. Intestinal bacteria living in the intestine are useful bacteria that protect human health and harmful bacteria that have a bad effect, and the health condition is controlled by the balance of both. Representative strains of useful bacteria are Lactobacillus sp. And Bifidobacterium sp., Which are lactic acid bacteria, and representative strains of harmful bacteria are Escherichia coli, food poisoning bacteria and Staphylococcus aureus. Get up quickly and produce carcinogens.

Lactobacillus is a bacterium (bacteria) that breaks down carbohydrates such as glucose or lactose and makes a lot of lactic acid. It is a bacterium that does not decompose proteins but does not decay, does not produce harmful substances, and has a beneficial effect. The lactic acid bacteria include Streptococcus sp., Pediococcus sp., And Leuconostoc sp. In addition to Lactobacillus and Bifidobacterium strains. Unlike other strains, lactic acid bacteria do not break down proteins to produce toxic substances such as ammonia, amines and hydrogen sulfide. Therefore, no matter how much lactic acid bacteria multiply in the intestine, it does not make a toxic substance, so it is thought to be a bacterium having a beneficial effect on the body.

The physiological activity of these lactic acid bacteria firstly, by reducing the pH of the intestine by producing an organic acid inhibits the growth of harmful bacteria in the intestine and maintains the normal intestinal flora. Secondly, it produces β-galactosidase, which favors people with lactose intolerance. Third, lowering the pH of the intestine has a direct effect on carcinogenesis (carcinogenesis), lactic acid, acetic acid inhibits the growth of decaying bacteria to reduce the carcinogens such as N-nitroso compounds, phenolic products exhibit anticancer activity. Fourthly, Bifidobacteria produces hydroxymethylglutarate and thus exhibits the effect of inhibiting HMG-CoA reductase and thus lowers cholesterol. Fifth, vitamins B ₁ , B ₂ , B ₆ , B ₁₂ , nicotinic acid and folic acid are synthesized. Sixth, diarrhea is prevented by suppressing the causative organism of diarrhea and normalizing the intestinal flora.

Enterobacteriaceae may be affected by the foods they eat, their living environment and stress, resulting in a loss of balance between harmful and beneficial bacteria. For example, Westerners who consume a lot of meat will have a lower intestinal beneficial bacterium, but Asians who consume more vegetables will have a lower incidence of colorectal cancer than Westerners due to a lower number of beneficial bacteria.

Two methods have been attempted to maintain lactic acid bacteria, which are beneficial bacteria in the intestinal flora, as dominant bacteria. First, the method of orally ingesting bifidus bacteria. Disadvantage of this method is that the intestinal bifidus bacillus exhibits the specificity of the host (ie, the person or animal in which the intestinal bacterium lives), so that the orally ingested bifidus bacterium cannot be excreted in the large intestine. Secondly, by orally ingesting bifidogenic factor, a substance selectively used by bifidus bacteria, if we ingest bifidus factor through food, we can easily grow lactic acid bacteria. Lactobacillus growth factors are selectively used by lactic acid bacteria and oligosaccharides are effective substances that are not absorbed in the digestive tract. In other words, it is a substance that inhibits the growth of harmful bacteria or pathogenic bacteria because it is used by intestinal useful bacteria including lactic acid bacteria in the colon and reaches the colon without digestion by digestive enzymes.

The lactic acid bacteria growth factor is, firstly, an amino sugar, and the bifidus factor containing the amino sugar includes β-ethyl-N-acetyl-D-glucosamine, N-acetylglucosamine and the like. Second, there are natural oligosaccharides such as lactose and inulin. Third, there are synthetic oligosaccharides, such as fructooligosaccharides.

Lactic acid bacteria catalyze reactions such as the generation of organic acids and do not cause rot reactions, and exhibit physiological activities such as inhibition of proliferation of harmful bacteria and immune enhancement. Therefore, lactic acid bacteria probiotics have been applied in clinical practice. Therefore, in order to use lactic acid bacteria preparations, they must be lyophilized and stored for cultivation and production as a product. However, in this process, lactic acid bacteria are easily killed. In particular, Bifidobacterium spp., Lactobacillus spp. And Clostridium butyricum strains are anaerobic strains, which are more stable than other lactic acid bacteria and are rapidly killed. Therefore, various stabilizers (protectors) have been developed.

The most important process in the production of lactic acid bacteria is lyophilization, which can be divided into freeze and dry. The former is a step of freezing the cells without causing injury (damage), and the latter is a step of subliming and drying the solvent (water) in the frozen sample under vacuum. In the course of these processes, crystalline forms of ice in lactic acid bacteria, salt concentrations, incomplete dehydration, complete dehydration, free radical accumulation, and infiltration pressure shock are the causes of lethal lactic acid bacteria. Therefore, attention should be paid to a number of factors in order to maintain stability for long-term preservation of live lactobacillus. In particular, protective substances (protectants) that can eliminate these causes are important for the stability of lactic acid bacteria preparations.

In the freezing process of lactic acid bacteria, the cooling rate is not easy to control, so the lactic acid bacteria are easily injured due to the difference between freezing process inside and outside the cells. Therefore, the protective substances used to control this process include glucose, sugar, lactose, skim milk, albumin and polyethylene glycol.

Drying of the bound water that remains unfrozen during the drying of frozen Lactobacillus bacteria is the main cause of damage to lactic acid bacteria. Therefore, glucose, sugar, amino acids (for example, glutamic acid, aspartic acid, citric acid, arginine, lysine) and the like are used to suppress the injury of lactic acid bacteria in this step.

Lyophilized biolactic acid bacteria are very dry, so if stored for a long time during this process, they are easily hygroscopic and easily killed. Therefore, glucose and glucosamine are used to suppress the injury of lactic acid bacteria during this storage process. In addition to this purpose gelatin, carboxymethyl cellulose, inositol, sorbitol and the like are used.

Korean Patent Registration No. 144130 discloses a ginseng polysaccharide having a specific structure, but only discloses that the ginseng polysaccharide has an immune enhancing effect, an anticancer effect, a cancer prevention effect, and a radiation protective effect. 2001-50186 discloses ginseng polysaccharides having a specific structure, but only the ginseng polysaccharides have been verified to have hematopoietic promotion, bone marrow defense, cancer cell killing immune cell production effect, cancer prevention effect and radiation sensitivity effect. Lactobacillus freeze-drying protection effect of the invention, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect and the inhibitory effect of the production of causative enzymes such as colon cancer are not mentioned at all.

It is an object of the present invention to provide a composition containing ginseng polysaccharides. Still another object of the present invention is to provide a composition containing ginseng polysaccharide having lactic acid bacteria freeze-drying protection effect, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect and production inhibitory effect of causative enzymes such as colon cancer. This use of the invention is a novel one completely different from the above known use.

The present invention provides a composition comprising at least one of ginseng, red ginseng or hot-treated ginseng polysaccharide 1-99.9% by weight based on the total weight of the composition.

The ginseng of the present invention includes all of Korean ginseng, American ginseng, Hwagi ginseng, Vietnamese ginseng, Himalayan ginseng, Jeonchisam and the like, and preferably Korean ginseng.

In addition, the present invention is the polysaccharide 40-50% hexasaccharides containing glucose or galactose, acidic polysaccharide 30-50% including galacturonic acid or rhamnose, or histidine, arginine, aspartic acid, threonine, serine or glutamic acid It provides a composition containing 3-6% protein comprising a.

Polysaccharides of ginseng, red ginseng and high-temperature ginseng contain 70-80% of starch and pectin-like components. Except for 20-30% starch, more than 20 polysaccharides such as Panaxan AU were isolated (Korea Ginseng and Tobacco Research Institute, Cheonil Printing Co., Daejeon, p75, 1994; Konno et al., Planta Med., 50, 434, 1984) . These polysaccharides are generally polymers with molecular weights of 10,000 Daltons or more, and constituent sugars contain 30-50% of acidic polysaccharides such as galacturonic acid and rhamnose and 40-50% of hexasaccharides such as glucose and galactose ( The Korean Ginseng Society, 14, 10, 1990), and others contain 3-6% of proteins composed of amino acids such as histidine, arginine, aspartic acid, threonine, serine, and glutamic acid.

The present invention also provides a composition containing the polysaccharide purified through the following process.

(a) about 1 to 20 times the amount (weight) of ginseng (water ginseng or white ginseng), red ginseng (ginseng dried at 95-100 ° C.) or hot-treated ginseng (ginseng dried at 100 ° C. or higher), preferably about 5 times Lower alcohols, such as fold to 10 times volume (L) of ethanol (50-100%), at an extraction temperature of 4 to 100 ° C., preferably 20 to 50 ° C., about 0.5 to 2 days, preferably 1 hour to A first step of extracting the extraction method such as extraction, reflux cooling extraction or ultrasonic extraction for one day by repeating the extraction once to five times, preferably two to three times;

(b) the residue remaining after removing the supernatant in the extract is about 1 to 20 times, preferably 10 times distilled water at 4 to 150 ℃, preferably 25 to 60 ℃ 1 to 24 hours, preferably 5 A second step of filtering the extract obtained by repeating an extraction method such as time extraction, reflux extraction or ultrasonic extraction once to five times, preferably two to three times;

(c) a third step of dialysis in the water flowing through the filtrate for 1 to 10 days, preferably 5 to 7 days;

(d) a fourth step of precipitating the dialysate with distilled water, lower alcohol or a mixed solvent having a mixing ratio of about 1: 0.1 to 1:10;

(e) a fifth step of suspending the precipitate of (d) in water and centrifuging again to obtain a supernatant;

The method for producing a polysaccharide from ginseng, red ginseng, or high-temperature treated ginseng proceeds in the order of (a), (b), (c), (d), and (e) (separation method 1). In some cases, all or part of the steps (c), (d) and (e) may be omitted (separation method 2).

In addition, the present invention provides a composition excellent in lactic acid bacteria freeze-drying protective effect, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect or production inhibitory effect of causative enzymes such as colon cancer.

The compositions of the present invention may further comprise suitable carriers, excipients and diluents according to conventional methods.

Carriers, excipients and diluents that may be included in the compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate and mineral oil.

The compositions of the present invention may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. .

The amount of the composition of the present invention may vary depending on the age, sex, and weight of the patient, but the amount of 0.1 to 500 mg / kg may be administered once to several times a day. In addition, the dosage may be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The composition of the present invention can be used in various formulations, such as lactic acid bacteria freeze-drying protective effect, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect or a drug having a production inhibitory effect of causative enzymes such as colon cancer. Examples of the food to which the ginseng polysaccharide composition can be added include various foods such as health foods such as various lactic acid bacteria, menopausal and senile foods, beauty foods, beverages, gums, teas, vitamin complexes, and the like. Since the composition has little toxicity and side effects, it is a drug that can be used safely even for long-term administration for the purpose of prevention. The amount of the composition that can be added to the food can generally be added in 0.01 to 15% by weight of the total food weight, the food health beverage composition is a ratio of 0.02 to 10 g, preferably 0.3 to 1 g based on 100 ml Can be added.

The liquid component of the health beverage composition of the present invention is not particularly limited, and may include various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. Examples of the natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol, erythritol and the like. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrate is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof. , Organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the composition of the present invention may include a natural fruit juice and a pulp for the production of fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of about 20% by weight or less per 100% by weight of the composition of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. However, these Examples and Experimental Examples are provided only for the purpose of illustration for better understanding of the present invention, and the scope and scope of the present invention are not limited by the following Examples.

Example 1

1-20 times (preferably 10 times) of ethanol was added to ginseng weight (g), and cold extraction was repeated twice at room temperature to obtain an extract and a residue. The remaining residue was heat treated at 60, 100 or 120 ° C. for 1-5 hours (preferably 3 hours), and then 1-20 times (preferably 10 times) of distilled water was added thereto, followed by 2 hours at 80 ° C. The extract obtained by extraction and filtration was dialyzed in running water for 7 days. The dialysis solution thus obtained was precipitated with 50% ethanol, and the precipitate was suspended in distilled water. The supernatant obtained by centrifugation as it was or again was lyophilized and used as a ginseng polysaccharide.

Example 2

The fresh ginseng is treated for 1-5 hours (preferably 3 hours) at 98 ° C and 70% of 1-20 times (preferably 10 times) to 1-24 hours (preferably 5 hours) dry red ginseng at 60 ° C. After extracting twice by cold extraction at room temperature with cold ethanol, 1-20 times (preferably 10 times) of distilled water was added to the remaining residue, and the resultant was extracted by filtration for 2 hours at 80 ° C and filtered. After precipitation with ethanol, the precipitate was suspended in distilled water, and the supernatant obtained by centrifugation as it was or lyophilized was used as red ginseng polysaccharide.

Example 3

Fine ginseng is treated for 1-5 hours (preferably 2 hours) at 120 ° C. under reduced pressure, and 1-20 times (preferably 10 times) of 70% ethanol is added to the high temperature dried ginseng dried at 60 ° C. for 5 hours at room temperature. The extract was extracted twice by cold extraction at, and 1-20 times (preferably 10 times) of distilled water was added to the remaining residue, followed by extraction at 80 ° C. for 2 hours, and the filtrate was precipitated with 50% ethanol. The precipitate was suspended in distilled water, and then the supernatant obtained by centrifugation as it was or lyophilized was used as a high-temperature ginseng polysaccharide.

Example 4

The fresh ginseng was treated for 3 hours at 98 ° C, and 1-20 times (preferably 10 times) of 70% ethanol was added to the red ginseng dried at 60 ° C for 5 hours, followed by cold extraction twice at room temperature. After adding 1-20 times (preferably 15 times) of distilled water, the supernatant obtained by centrifugation of the extract obtained by extracting and filtering at 4 ° C. for 15 hours was used as red ginseng polysaccharide.

Determination of Sugar Content of Isolated Ginseng, Red Ginseng, and High-Temperature Ginseng Polysaccharides

The sugar content of jinsan to polysaccharides isolated from these ginsengs was measured by the Antron-Sulfate method. A solution of separated ginseng polysaccharides in distilled water and a double dilution series solution at a concentration of 1 mg / mL of the sample was shaken on ice with 3 ml of anthrone-sulfuric acid reagent, which was stored at 0.25 ° C at 4 ° C, for 10 minutes. The content was measured at 620 nm. At this time, glucose was used as a standard product and was prepared from a calibration curve.

Acidic Polysaccharide Contents of Isolated Ginseng, Red Ginseng, and High-Temperature Ginseng Polysaccharides

The isolated polysaccharide was measured by measuring the uronic acid content. Separate the solution of polysaccharide sample from 10 mg / mL polysaccharide solution in distilled water and dilute it twice with 0.4 ml and 0.0125 M sodium sulfate-borate, respectively, and shake well for 100 minutes. After adding 0.04 ml of 0.15% m-hydroxybiphenyl solution on ice, shaking well, lowering the temperature for another 15 minutes, cooling at room temperature, and calculating the content by measuring absorbance at 520 mm. A calibration curve was prepared using galacturonic acid as a standard product.

The red ginseng polysaccharide solution was identified using a Sephadex column (Sephadex G-75, 1.6 × 50 cm, Pharmacia) to confirm its elution pattern. The column was eluted with distilled water and the flow rate was 10 mL / h. The eluted fractions were fractionated by 0.5 ml. Molecular weight markers measured aldolase, bovine serum albumin, ribonuclease A at 280 nm, and the polysaccharides eluted from the column were absorbed at 540 nm after color development by carbazole reaction. . The elution pattern of the red ginseng polysaccharide was as shown in Figure 1, the molecular weight of the red ginseng polysaccharide was about 1-10 million daltons, the most was about 4-70,000 daltons. Ginseng polysaccharides and high-temperature treated ginseng polysaccharides showed similar results.

As a result of the measurement by the above experimental method, sugar, acidic polysaccharide and protein content are shown in Table 1 below.

Hexose Sugar Content (%) Acidic polysaccharide content (%) Protein content (%) Molecular weight (kilodaltons) 60 ° C treated ginseng of Example 1 > 38.2 8.9 1.1 10-> 100 100 ° C treated ginseng of Example 1 > 34.5 26.6 1.5 10-100 120 ° C treated ginseng of Example 1 > 23.2 24.4 1.3 10-100 Example 2 > 35.2 19.9 2.1 10-100 Example 3 > 43.2 19.2 2.8 10-100 Example 4 > 42.1 13.4 2.9 10-100

Lactic acid bacteria stability effect

Experimental Example 1

The strain LB Bifidobacterium breve K-110 (KCCM 10097), Lactobacillus acidophilus (KCTC 3168), Streptococcus faecium (KCTC 3077) (Tryptic Soy Broth) Bacteria obtained by culturing in medium or synthetic medium (containing edible yeast extract, vitamin C, dextrose) are collected by centrifugation and washed twice with physiological saline. This strain was suspended by adding 10% of skim milk, glucose, ginseng (white ginseng) polysaccharide, red ginseng polysaccharide, or high-temperature treated ginseng polysaccharide, quenched with liquefied nitrogen for 10 minutes, and dried by lyophilizer (Japan, Eyler Co.). The number of bacteria was measured, and then the stability of the strain was measured while storing at 4 ℃ and 25 ℃. On the other hand, lactic acid bacteria of the present invention is not limited to specific lactic acid bacteria (KCCM (Korean Culture Center of Microorganisms) Korea Microorganism Conservation Center; KCTC (Korean Collection for Type Cultures) Korea Institute of Biotechnology Gene Bank).

Survival rate (%) = (number of cells survived / number of cells used first) x 100

Stabilization Effect of Ginseng Polysaccharides on Lyophilization of Lactic Acid Bacteria Survival rate (%) Bifidobacterium breb K-110  Lactobacillus asidophilus  Streptococcus Passium none <1 <1 <1 Ginseng Separation Polysaccharide (Separation Method 1) 8.5 8.7 9.1 Ginseng Separation Polysaccharide (Separation Method 2) 8.3 8.9 9.2 Red Ginseng Isolation Polysaccharide (Isolation Method 1) 9.5 10.2 10.5 Red Ginseng Separation Polysaccharide (Separation Method 2) 9.0 9.8 10.5 High Temperature Ginseng Separation Polysaccharide (Separation Method 1) 9.2 8.9 11.0 High Temperature Ginseng Separation Polysaccharide (Separation Method 2) 8.2 8.5 10.2 Scheme milk 10.2 10.0 10.3 Lactose 9.5 8.9 9.3

Considering that lactose is a sugar that causes lactose intolerance for many people, especially Asians including Koreans, ginseng polysaccharides are excellent as lyophilized protective agents.

Stabilizing Effect of Ginseng Polysaccharides during Storage of Lactic Acid Bacteria at 4 ℃ for 1 Month Survival rate (%) Bifidobacterium breb K-110 Lactobacillus asidophilus Streptococcus Passium none <1 <1 <1 Ginseng Separation Polysaccharide (Separation Method 1) 75 86 91 Ginseng Separation Polysaccharide (Separation Method 2) 75 76 92 Red Ginseng Isolation Polysaccharide (Isolation Method 1) 99 91 95 Red Ginseng Separation Polysaccharide (Separation Method 2) 98 86 98 High Temperature Ginseng Separation Polysaccharide (Separation Method 1) 87 88 96 High Temperature Ginseng Separation Polysaccharide (Separation Method 2) 78 85 95 Scheme milk 38 45 65

Lactobacillus prepared by adding different lactic acid bacterium stabilizing agents was stored at 4 ℃, and ginseng polysaccharides, red ginseng polysaccharides, and high-temperature treated ginseng polysaccharides showed better stabilizing effects than current lactose stabilizers.

Stabilizing Effect of Ginseng Polysaccharides during Storage of Lactic Acid Bacteria at 4 ℃ for 1 Month Survival rate (%) Bifidobacterium breb K-110 Lactobacillus asidophilus Streptococcus Passium none <1 <1 <1 Scheme Milk + Ginseng Separation Polysaccharide (Separation Method 1) 78 82 88 Scheme Milk + Ginseng Separation Polysaccharide (Separation Method 2) 68 79 89 Scheme milk + red ginseng isolated polysaccharide (Separation method 1) 81 91 93 Scheme milk + red ginseng isolated polysaccharide (separation method 2) 79 82 93 Scheme milk + high temperature ginseng separation polysaccharide (separation method 1) 83 85 89 Scheme milk + high-temperature ginseng separation polysaccharide (separation method 2) 72 79 92 Scheme milk 38 45 65

Lactobacillus prepared by adding different lactic acid stabilizer and lyophilized protector at the same time was measured at 4 ° C and the number of viable cells was also better than lactose stabilizer. Ginseng polysaccharides, red ginseng polysaccharide, The treated ginseng polysaccharides were found to contribute more significantly.

Stabilizing Effect of Ginseng Polysaccharides during Storage of Lactic Acid Bacteria at 25 ℃ for 1 Month Survival rate (%) Bifidobacterium breb K-110 Lactobacillus asidophilus Streptococcus Passium none <1 <1 <1 Ginseng Separation Polysaccharide (Separation Method 1) 13 14 16 Ginseng Separation Polysaccharide (Separation Method 2) 14 12 12 Red Ginseng Isolation Polysaccharide (Isolation Method 1) 15 18 21 Red Ginseng Separation Polysaccharide (Separation Method 2) 14 15 19 High Temperature Ginseng Separation Polysaccharide (Separation Method 1) 18 14 26 High Temperature Ginseng Separation Polysaccharide (Separation Method 2) 21 16 22 Scheme milk 17 12 21

Ginseng polysaccharides, red ginseng polysaccharides, and high-temperature treated ginseng polysaccharides showed similar or superior stabilization effects to lactose, a good lactic acid bacteria stabilizer, when the number of live cells was stored at 25 ° C.

Lactobacillus multiplication effect

Experimental Example 2

Ginseng extract polysaccharides and red ginseng extract polysaccharides were added to the GAM medium to 0.25, 0.5%, respectively, and autoclave sterilized and lactulose (lactulose), known as a lactic acid bacteria growth factor, was used as a control material. Bifidobacterium Breb K-110 (KCCM 10097), Bifidobacterium longum H-1 (KCCM 10493), Lactobacillus mesenteroides ( Lactobacillus mesenteroides ) (Kyung Hee University Pharmacy Microbiology Class) were transplanted to measure pH and the number of lactic acid bacteria.

Proliferation Rate (%) Bifidobacterium breb K-110 Bifidobacterium longgum H-1 Lactobacillus asidophilus Streptococcus Passium Lactobacillus mecenteroides GAM (-dextrose) medium 100 100 100 100 100 Ginseng Separation Polysaccharide (Separation Method 1) 180 132 122 210 154 Ginseng Separation Polysaccharide (Separation Method 2) 124 170 121 192 181 Red Ginseng Isolation Polysaccharide (Isolation Method 1) 132 189 160 182 190 Red Ginseng Separation Polysaccharide (Separation Method 2) 190 180 150 213 178 High Temperature Ginseng Separation Polysaccharide (Separation Method 1) 130 134 152 192 170 High Temperature Ginseng Separation Polysaccharide (Separation Method 2) 140 154 250 205 166 Lactulose 230 238 350 240 230

Lactic acid bacteria proliferation effect of polysaccharides of ginseng, red ginseng, and high-temperature treated ginseng was excellent.

Growth inhibitory effect of harmful bacteria

Experimental Example 3: Effect on Enterobacteriaceae

Dilute human enterobacteriaceae to 10 ⁵ and transplant it into culture medium made by adding ginseng polysaccharide or red ginseng polysaccharide to 5 ml of GAM (-dextrose) medium, and cultured for 24 hours, and then colon cancer and soy sauce produced by the strain Enzyme activity of β-glucuronidase and tryptophanase, the causative enzymes of the sun, was measured.

 β-glucuronidase enzyme activity was reacted for 1 hour at 37 ° C by adding 0.02 ml of 10 mM p-nitrophenyl-β-D-glucuronide and 0.1 ml of enzyme solution to 0.38 ml of 0.1 M phosphate buffer (pH 7.0). The reaction was terminated by adding 0.5 mL of 0.5 N NaOH, 1 mL of distilled water was added, followed by centrifugation (2000 × g, 20 min), and the absorbance was measured at 405 nm for the supernatant.

Tryptophanase enzyme activity was obtained by adding 0.2 ml of 0.02 M tryptophan and 0.1 ml of enzyme solution by adding 30 ml of a complete reaction mixture (0.1 M bisine pH 8.0), 4% pyridoxal 5-phosphate, 20% calf serum albumin) After the reaction was completed, 2 ml of a colorant (14.7 g of p-dimethylaminobenzaldehyde, 948 ml of 95% ethanol, 52 ml of CH ₂ SO ₄ ) was added to terminate the reaction, followed by centrifugation (2000 × g, 20 minutes), followed by a supernatant solution. Absorbance was measured at 550 nm.

Table 7 below shows the inhibition rate of colon cancer and hepatic inducing enzymes when E. coli HGU-3 (β-glucuronidase-producing bacteria) and lactic acid bacteria were mixed in culture.

Polysaccharide added to GAM (-dextrose) medium Inhibition Rate (%) β-glucuronidase Tryptophanase none 0 0 Ginseng Separation Polysaccharide (Separation Method 1) 52 89 Ginseng Separation Polysaccharide (Separation Method 2) 58 45 Red Ginseng Isolation Polysaccharide (Isolation Method 1) 67 75 Red Ginseng Separation Polysaccharide (Separation Method 2) 75 72 High Temperature Ginseng Separation Polysaccharide (Separation Method 1) 70 81 High Temperature Ginseng Separation Polysaccharide (Separation Method 2) 69 68 Lactulose 85 62

Polysaccharides of ginseng or red ginseng showed an effect of significantly inhibiting the production of β-glucuronidase and tryptophanase produced by enterobacteriaceae known as carcinogen.

As described above, the composition containing the ginseng polysaccharide of the present invention is excellent in the lactic acid bacteria lyophilization protective effect, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect or production inhibitory effect of causative enzymes such as colon cancer.

Claims (8)

A composition containing 1-99.9% by weight of any one or more of the polysaccharide of ginseng, red ginseng or hot-treated ginseng based on the total weight of the composition. 2. The polysaccharide according to claim 1, wherein the polysaccharide is 40-50% hexasaccharide with glucose or galactose, acidic polysaccharide 30-50% with galacturonic acid or rhamnose, or histidine, arginine, aspartic acid, threonine, serine or glutamic acid. A composition containing 3-6% protein comprising a. The food composition according to claim 1 or 2, having a lyophilized protective effect for probiotic formulation of lactic acid bacteria. The food composition according to claim 1 or 2, having a storage stabilizing effect for probiotic formulation of lactic acid bacteria. The food composition of Claim 1 or 2 which has a lactic acid bacteria growth effect. The pharmaceutical composition according to claim 1 or 2, for treating colorectal cancer or liver injury. The composition of claim 1 or 2, wherein the polysaccharide is purified through the following process: (a) Extracting methods such as ginseng, red ginseng or hot-treated ginseng with a lower alcohol such as 1 to 20 times the volume of ethanol for 0.5 hours to 2 days at an extraction temperature of 4 to 100 ° C., reflux cooling extraction or ultrasonic extraction A first step of extracting by repeating 1 to 5 times; And (b) an extract obtained by removing the supernatant from the extract and repeating an extraction method such as extraction, reflux extraction or ultrasonic extraction at 1 to 24 hours at 4 to 150 ° C. with distilled water of 1 to 20 times 1 to 5 times. Filtering the second step. The composition of claim 1 or 2, wherein the polysaccharide is purified through the following process: (a) Extracting methods such as ginseng, red ginseng or hot-treated ginseng with a lower alcohol such as 1 to 20 times the volume of ethanol for 0.5 hours to 2 days at an extraction temperature of 4 to 100 ° C., reflux cooling extraction or ultrasonic extraction A first step of extracting by repeating 1 to 5 times; (b) an extract obtained by removing the supernatant from the extract and repeating an extraction method such as extraction, reflux extraction or ultrasonic extraction at 1 to 24 hours at 4 to 150 ° C. with distilled water of 1 to 20 times 1 to 5 times. Filtering the second step; (c) a third step of dialysis of the filtrate in running water for 1 to 10 days; (d) a fourth step of precipitating the dialysate with distilled water, lower alcohol or a mixed solvent having a mixing ratio of 1: 0.1, 1: 10; And (e) a fifth step of suspending the precipitate of (d) in water and centrifuging again to obtain a supernatant, provided that any one or more of steps (c) to (e) can be omitted).

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